Identification of novel cancer-specific antigens and application for cellular imunotherapy of hematological malignancies

新型癌症特异性抗原的鉴定及其在血液恶性肿瘤细胞免疫治疗中的应用

• 批准号: 13470206
• 负责人: YASUKAWA Masaki
• 金额: $ 8.83万
• 依托单位: Ehime University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470206/
• 关键词: Cancer antigens; Immunotherapy; Leukemia; Cytotoxic T lymphocytes; HLA; WT1; Telomerase; HLA-A24; 肺がん; 自殺遺伝子

To identify the novel cancer-associated antigens and to develop the novel immunotherapy for hematological malignancies, this study was performed. The data obtained from the series of experiments are as follows.1) A WT1-specific, HLA-A24-restricted CTL clone (designated TAK-1) exhibited cytotoxicity against lung cancer cell lines bearing HLA-A24 but did not lyse cells lacking this HLA. Adoptive transfer of TAK-1 into nude mice that had been engrafted with an HLA-A24-positive lung cancer cell line resulted in inhibition of the cancer cell growth and prolonged survival. These findings strongly suggest that WT1 is a universal tumor-associated antigen and that WT1 -targeting immunotherapy offers a potentially effective treatment option for lung cancer as well as leukemia.2) A novel WT1-derived peptide-specific CD8^+ CTL line, designated NIM-1 was established. NIM-1 lysed HLA-A24-positive leukaemia cells, but not HLA-A24 negative leukaemia cells or normal cells.3) Immature dendritic cells (D … More Cs) were loaded with leukemia cells with t(6 ; 9) or t(9 ; 22) and then cocultured with the dek-can fusion peptide-specific or the bcr-abl fusion peptide-specific CD4^+ T-lymphocyte clone. The dek-can peptide-specific and bcr-abl peptide-specific CD4^+ T-lymphocyte clones produced interferon-γ(IFN-γ) when they were cocultured with HLA-DR-matched but not with mismatched DCs which had been loaded with apoptotic as well as necrotic leukemia cells with t(6 ; 9) and t(9 ; 22), respectively. These data indicate that the acute myelogenous leukemia-associated fusion protein, dek-can, and chronic myelogenous leukemia-associated fusion protein, bcr-abl, are both processed and presented by DCs to the fusion peptide-specific CD4^+ T lymphocytes.4) In order to clarify the roles of perforin in antigen-specific cytotoxicity mediated by human CD4^+ CTLs, antigen-specific human CD4^+ T-lymphocyte clones were established from a patient with hereditary perforin deficiency and their cytotoxic activities were investigated. The data demonstrated that perforin-negative CD4^+ CTLs can exert cytotoxicity against Fas-sensitive target cells ; however, perforin plays essential roles in antigen-specific cytotoxicity mediated by human CD4^+ as well as CD8^+ CTLs. Less

为了鉴定新的癌症相关抗原和开发新的免疫治疗血液系统恶性肿瘤，本研究进行。从一系列实验中得到的数据如下。1) wt1特异性，HLA- a24限制性CTL克隆（指定TAK-1）对携带HLA- a24的肺癌细胞系显示细胞毒性，但不裂解缺乏该HLA的细胞。将TAK-1过继转移到移植了hla - a24阳性肺癌细胞系的裸鼠体内，可抑制癌细胞生长，延长存活时间。这些发现有力地表明，WT1是一种普遍的肿瘤相关抗原，WT1靶向免疫疗法为肺癌和白血病提供了一种潜在的有效治疗选择。2)建立了一种新的wt1衍生肽特异性CD8^+ CTL，命名为nim1。nim1能溶解HLA-A24阳性白血病细胞，但不能溶解HLA-A24阴性白血病细胞或正常细胞。3)未成熟树突状细胞（D…More Cs）装载t（6; 9）或t（9; 22）的白血病细胞，然后与dek-can融合肽特异性或bcr-abl融合肽特异性CD4^+ t淋巴细胞克隆共培养。dek-can肽特异性和bcr-abl肽特异性CD4^+ t淋巴细胞克隆在与hla - dr匹配的dc共培养时产生干扰素-γ(IFN-γ)，而与t（6; 9）和t（9; 22）分别装载凋亡和坏死白血病细胞的错配dc共培养时则不产生干扰素-γ。这些数据表明，急性髓性白血病相关融合蛋白dek-can和慢性髓性白血病相关融合蛋白bcr-abl都被dc加工并提呈给融合肽特异性CD4^+ T淋巴细胞。4)为了明确穿孔素在人CD4^+ ctl介导的抗原特异性细胞毒性中的作用，我们建立了遗传性穿孔素缺乏症患者的抗原特异性CD4^+ t淋巴细胞克隆，并对其细胞毒活性进行了研究。数据表明，穿孔素阴性CD4^+ ctl可对fas敏感靶细胞发挥细胞毒性；然而，穿孔素在人类CD4^+和CD8^+ ctl介导的抗原特异性细胞毒性中起着重要作用。少

项目成果

Azuma, T., et al.: "Identification of a novel WT1-derived peptide which induces HLA-A24-restricted anti-leukaemia cytotoxic T lymphocytes"British Journal of Haematology. 116. 601-603 (2002)

Azuma，T.，等人：“诱导 HLA-A24 限制性抗白血病细胞毒性 T 淋巴细胞的新型 WT1 衍生肽的鉴定”英国血液学杂志。

Arai, J., et al.: "Identification of human telomerase reverse transcriptase-derived peptides which induce HLA-A24-restiricted antileukemia CTLs"Blood. 97. 2903-2907 (2001)

Arai, J. 等人：“诱导 HLA-A24 限制性抗白血病 CTL 的人端粒酶逆转录酶衍生肽的鉴定”血液。

Makita, M., Yasukawa, M.et al.: "Antilung cancer effect of WT1-specific cytotoxic T lymphocytes"Clinical Cancer Research. 8. 2626-2613 (2002)

Makita, M., Yasukawa, M.等人：“WT1特异性细胞毒性T淋巴细胞的抗肺癌作用”临床癌症研究。

Kakimoto, M., Hasegawa, A., Fujita, S. and Yasukawa, M.: "Phenotypic and functional alterations of dendritic cells induced by human herpesvirus 6 infection"J.Virol.. 76. 10338-10345 (2002)

Kakimoto, M.、Hasekawa, A.、Fujita, S. 和 Yasukawa, M.：“人疱疹病毒 6 感染诱导的树突状细胞的表型和功能改变”J.Virol.. 76. 10338-10345 (2002)

Hasegawa, A., et al.: "Transcriptional down-regulation of CXCR4 induced by impaired association of YY1 with c-Myc in HHV-6-infected cells"Journal of Immunology. 166. 1125-1131 (2001)

Hasekawa, A. 等人：“HHV-6 感染细胞中 YY1 与 c-Myc 的关联受损导致 CXCR4 转录下调”《免疫学杂志》。